1. Field of the Invention
The present invention relates to pharmaceutical compositions, methods and kits that provide for the early diagnosis and treatment of lung cancer. More particularly, the present invention relates to pharmaceutical compositions containing uteroglobin for treating or inhibiting metastasis of lung epithelial tumor cells and methods of using the same to treat or inhibit metastasis of lung epithelial tumor cells. The present invention also relates to methods and kits for early diagnosis of metastatic lung cancer by assaying for uteroglobin and comparing the results against control cells.
2. Description of the Prior Art
Cancers develop from uncontrolled multiplication of cells. All cancers are life threatening, and lung cancer remains the major cause of cancer death among both males and females.
There are four types of lung cancer found in humans: squamous, adeno, small cell, and large cell. Each tumor expresses specific differentiation features or surface phenotype determinants, all of which distinguish these cells form normal cells. The development of monoclonal antibody diagnostic techniques has greatly enhanced the production of reagents capable of differentiating normal cells from cancer cells and differentiating between cancer cell types. However, none of these markers have been able to provide information concerning when a tumor cell or cells will become metastatic.
The major cause of morbidity and mortality for cancer in humans is metastatic disease. As a consequence, there has been much interest in the mechanisms involved in invasion of cells and metastasis. Several enzyme systems have been implicated in the metastatic process: metalloproteinases, cysteine proteases, and serine proteases. Yagel, S. A. et al., 49 Cancer Research 3553 (1989), Dickson, R. B., 41 J. Steroid Biochem. Molec. Biol. 389 (1992) and Zucker S. et al., 45 Cancer Research 6168 (1985). Inhibitors of metalloproteinases, especially of the collagenases, have been the focus of intense study. DeClerck A. et al., 52 Cancer Research 701 (1992).
However, metastatic cancers have proven to be particularly difficult to treat. These cancers pose the highest risk to patients and, for optimal prognosis, often must be treated by aggressive methods that present increased risks of deleterious side-effects. Most treatments now available have severe toxic side effects to the human body such as nausea, vomiting, hair loss and fatigue.
Therefore, there is a great need for methods that accurately distinguish those tumors that are likely to metastasize from those that are unlikely to do so. Furthermore, since currently available methods of treating metastatic cancers often are inadequate, there also is a clear need for improved anti-metastatic agents and methods to treat metastatic cancers, and metastatic lung cancers in particular.
Metastatic cancers originate from a primary tumor. Metastasis of the primary tumor produces secondary tumors and disseminated cancer. It is well known that both primary and secondary tumors shed large numbers of cells.
The shed cells can spread through the body. For instance, a primary tumor may damage the surrounding lymph or circulatory vessels, allowing entry of shed cells into the lymph or circulatory systems, and hastening their spread in the body. Moreover, shedding of cells by cancerous tumors increases during surgery and radiotherapy.
Most shed cells do not form new tumors. To do so such cells must surmount a series of physical and physiological barriers. In fact, a series of distinct events must occur for metastasis to occur.
The primary tumor physically must (i) invade interstitial space of the primary tissue. In particular, it must (ii) penetrate the basement membrane of the tissue. For most metastases, the tumor must damage the endothelial cell wall of lymphatic or vascular vessels to provide access to shed cells. Cells that enter the lymph or blood must (iii) survive hemodynamic stress and host defenses in the circulation and, furthermore, (iv) the cells must lodge at a new site in the circulatory system, a process that apparently involves aggregated platelets. A cell then must (v) extravasate out of the vessel into the interstitial space. Finally, it must (vi) invade the interstitial space of the secondary organ and proliferate in the new location. Although the process of metastasis is physiologically complex, the overall pattern of metastasis is general to many types of cancers including lung cancers.
The metastatic process also clearly involves complex intracellular mechanisms that alter cancerous cells and their interactions with surrounding cells and tissues. Currently, it is thought that proliferation of many cancerous cells depends upon specific ligand-receptor interactions. Thus far, however, it has not been possible to develop a therapy that prevents or effectively inhibits metastasis of metastatic cancers.
The complexity of the processes involved in metastasis, and the lack of understanding of underlying molecular mechanisms, have made it particularly difficult in some cases to distinguish tumors that are likely to metastasize from those that are unlikely to do so.
The inability to discern the metastatic potential of tumors precludes accurate prognosis and leads, inevitably, to the therapeutic intervention that either is too aggressive or insufficiently aggressive. Furthermore, for all types of cancers it has been difficult or impossible thus far to develop treatments that inhibit or prevent the spread of metastatic tumors. Clearly, there remains a great need for methods to accurately determine the metastatic potential of tumors and for effective anti-metastatic compositions and methods.
The development of monoclonal antibody techniques for differentiating normal cells from cancer cells and differentiating one type of cancer cell from another has greatly enhanced lung cancer detection but does not yield information regarding metastatic potential of a neoplastic, dysplastic, or tumor cell.
Mulshine et al., U.S. Pat. No. 5,455,159, discloses early diagnosis of lung cancer by using monoclonal antibodies to detect cells that express antigens whose increased levels correlates with the development of lung cancer.
Mulshine et al., U.S. Pat. No. 4,569,788, discloses monoclonal antibodies which can be used to detect non-small cell lung cancer and distinguish non-small cell cancer from other cancers and normal cells.
Hirohashi et al., U.S. Pat. No. 4,683,200, discloses IgM class monoclonal antibody which is reactive with human lung cancers.
Loor et al., U.S. Pat. No. 4,690,890, discloses a process for detecting two antigens using an immunometric dual sandwich assay.
Tanswell et al., U.S. Pat. No. 4,624,930, discloses a process for detecting polyvalent antigens using a three receptor reaction.
However, none of the prior art techniques utilize uteroglobin or antibodies which react with the same. None of the prior art techniques disclose the use of proteins which are down-regulated or which exhibit decreased expression of the normal protein during neoplastic development. In contrast, the prior art has generally relied upon the up-regulation of cellular proteins to support the detection of these tumor markers. Further, the prior art does not disclose aberrantly processed proteins leading to aberrant protein structure and function, nor does the prior art provide for the ability to inhibit the metastatic process by administering the missing normal protein, thus re-creating positive feedback mechanisms within the cellular machinery.
The present invention relates to pharmaceutical compositions, methods and kits that provide for the early diagnosis and treatment of lung cancer. More particularly, the present invention relates to pharmaceutical compositions containing uteroglobin protein of SEQ ID NO. 1 for treating or inhibiting metastasis of lung epithelial tumor cells and methods of using the same to treat or inhibit metastasis of lung epithelial tumor cells. The present invention also relates to methods and kits for early diagnosis of metastatic lung cancer by assaying for uteroglobin protein of SEQ ID NO. 1 and comparing the results against control cells.
Accordingly, the present invention provides a method for treatment of metastatic lung cancer, comprising administering a therapeutically effective amount of an inhibitor of phospholipase A2 to an animal.
The present invention also provides a pharmaceutical composition for treatment of metastatic lung cancer in an animal, comprising a therapeutically effective amount of an inhibitor of phospholipase A2 and a pharmaceutically acceptable carrier.
The present invention is also directed to a method for inhibiting metastasis of lung epithelial tumor cells, comprising administering a therapeutically effective amount of an inhibitor of phospholipase A2 to an animal.
The present invention is further directed to a pharmaceutical composition for inhibiting metastasis of lung epithelial tumor cells in an animal, comprising a therapeutically effective amount of an inhibitor of phospholipase A2 and a pharmaceutically acceptable carrier.
The present invention additionally provides for a method for inhibiting invasion of lung epithelial tumor cells, comprising administering a therapeutically effective amount of an inhibitor of phospholipase A2 to an animal.
A pharmaceutical composition for inhibiting invasion of lung epithelial tumor cells in an animal is also provided and comprises a therapeutically effective amount of an inhibitor of phospholipase A2 and a pharmaceutically acceptable carrier.
In addition to methods and compositions for treating metastatic lung cancer cells, the present invention also relates to diagnostic methods and kits.
One preferred method is directed to a method for detecting or identifying metastatic lung cancer, comprising: comparing an amount of an inhibitor of phospholipase A2 in a sample of lung epithelial tissue to at least one reference which correlates to the amount of inhibitor of phospholipase A2 in normal lung epithelial tissue, or to metastatic lung cancer, whereby differential amounts of the inhibitor of phospholipase A2 between the sample of lung epithelial tissue and the reference, detects or identifies metastatic lung cancer.
Another preferred method includes detecting or identifying metastatic lung cancer, comprising: assaying for inhibitor of phospholipase A2 in a sample of lung epithelial tissue; and comparing the amount of the inhibitor in the sample to a reference which correlates to the amount of the inhibitor in normal lung epithelial cells or metastatic lung epithelial cells, wherein differential amounts of the inhibitor of between the sample and the reference detects or identifies metastatic lung cancer.
An additional preferred method contemplates detecting or identifying a pathological condition of lung epithelial tissue, comprising: assaying for inhibitor of phospholipase A2 in a sample of lung epithelial tissue; and comparing the amount of the inhibitor in the sample to a reference which correlates to the amount of the inhibitor in normal lung epithelial tissue or metastatic lung epithelial cells, wherein differential amounts of the inhibitor between the sample and the reference detects or identifies a pathological condition of lung tissue.
A further preferred embodiment of the present invention includes a method for detecting or identifying an inhibitor of phospholipase A2 in a sample of lung epithelial tissue, comprising: assaying for an inhibitor of phospholipase A2 in a sample of sample of lung epithelial.
Another particularly preferred embodiment of the invention relates to the detection of aberrant uteroglobin protein of SEQ ID NO. 1. Surprisingly, an aberrant form of native uteroglobin protein of SEQ ID NO. 1 is expressed and directly correlates to the decrease in native or normal uteroglobin protein of SEQ ID NO. 1 as metastasis progresses. Accordingly, one aspect of the present invention involves not only the detection of the loss of normal uteroglobin protein of SEQ ID NO. 1 as a gauge of metastatic or invasive activity, but also the increase in the level of aberrant uteroglobin protein of SEQ ID NO. 1. This increase provides a positive detection whereas the loss of normal uteroglobin protein of SEQ ID NO. 1 relies upon the absence. Furthermore, this aberrant form of uteroglobin protein of SEQ ID NO. 1 is secreted in sputum and bronchial fluid, as well as other related bodily fluids such as respiratory fluids and the like where such proteins are found. Accordingly, a method and kit for detection of an aberrant form of uteroglobin protein of SEQ ID NO. 1 as a direct indicator of metastasis and invasiveness is provided.
Preferred kits of the present invention include a kit for treatment of lung cancer, comprising a therapeutically effective amount of an inhibitor of phospholipase A2 in a pharmaceutically acceptable carrier and a device for delivery of the inhibitor to the lung cancer, wherein the inhibitor, the carrier and the device are packaged in a container.
An additional preferred kit contemplates inhibiting metastasis of lung epithelial tumor cells, comprising a therapeutically effective amount of an inhibitor of phospholipase A2 in a pharmaceutically acceptable carrier and a device for delivery of the inhibitor to the lung epithelial tumor cells, wherein the inhibitor, the carrier and the device are packaged in a container.
A further kit of the present invention is a kit for inhibiting invasion of lung epithelial tumor cells, comprising a therapeutically effective amount of an inhibitor of phospholipase A2 in a pharmaceutically acceptable carrier and a device for delivery of the inhibitor to the lung epithelial tumor cells, wherein the inhibitor, the carrier and the device are packaged in a container.
The present invention also provides a kit for detecting or identifying metastatic lung cancer, comprising: a) means for collecting a sample of lung epithelial cells; b) means for detecting an inhibitor of phospholipase A2 in the sample of lung epithelial cells; and c) at least one reference which correlates to the amount of inhibitor of phospholipase A2 in normal lung epithelial cells or in metastatic lung cancer cells, wherein differential amounts of the inhibitor of phospholipase A2 between the sample and the reference detects or identifies metastatic lung cancer.
Another preferred kit is a kit for detecting or identifying a pathological condition of lung epithelial cells, comprising: a) means for collecting a sample of lung epithelial cells; b) means for detecting an inhibitor of phospholipase A2 in the sample of lung epithelial cells; and c) at least one reference which correlates to the amount of inhibitor of phospholipase A2 in normal lung epithelial cells or in metastatic lung cancer cells, wherein differential amounts of the inhibitor of phospholipase A2 between the sample and the reference detects or identifies a pathological condition of the lung epithelial cells.
A further kit of the present invention involves a kit for detecting or identifying an inhibitor of phospholipase A2 in a sample of lung epithelial cells, comprising: a) means for collecting a sample of lung epithelial cells; and b) means for detecting an inhibitor of phospholipase A2 in the sample of lung epithelial cells.